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Method for measuring glucose oxidase without interference of blood lipid

A technology of glucose oxidase and a determination method, which is applied in the field of determination containing enzymes, can solve problems such as endogenous lipid blood interference, and achieve the effect of high accuracy

Inactive Publication Date: 2018-04-06
TIANJIN BAODI HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In order to solve the problem of endogenous lipemia interference in the prior art serum glucose assay method, the present invention provides an economical, convenient, and highly accurate glucose oxidase assay method that can effectively eliminate endogenous lipemia interference

Method used

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  • Method for measuring glucose oxidase without interference of blood lipid
  • Method for measuring glucose oxidase without interference of blood lipid
  • Method for measuring glucose oxidase without interference of blood lipid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Composition of reagents:

[0025] a. Reagent I:

[0026] Contains NAD in 0.1mol phosphate buffer per liter + 6.0mmol, lipoprotein lipase 300U, glycerol dehydrogenase 750U, Triton X-1000.15g, magnesium sulfate 2.0mmol / L, mutarotase 1.5KU, Proclin-300 preservative 200μl, among them, Triton X-100 is lipoprotein The dissociating agent, lipoprotein lipase, and glycerol dehydrogenase are triglyceride decomposing enzymes.

[0027] b. Reagent II:

[0028] Each liter of 0.1mol phosphate buffer contains ascorbate oxidase 8.0KU, glucose oxidase 15KU, peroxidase 15KU, 4-aminoantipyrine 0.5mmol, 2,4-dichlorophenol 0.5mmol / L, Proclin -300200 μl.

[0029] Among them, magnesium sulfate is an activator of glycerol dehydrogenase, Triton X-100 is a lipoprotein dissociation agent, and Proclin-300 is a preservative.

[0030] c. Standard solution: 5.55mmol / L glucose aqueous solution.

[0031] The volume ratio of the reactants in the above determination is: sample: reagent I: reagent II...

Embodiment 2

[0033] Measurement procedure

[0034] Research objects 204 cases of inpatients and outpatients took 2.0mL fasting blood samples, centrifuged at low speed for 10 minutes, and separated serum. In the lipidemia group, glucose was measured by the method of the present invention and the glucose measurement method of Leadman Biochemical Co., Ltd. after ultracentrifugation, and the real-time response curve was observed.

[0035] The detection effect of the present invention is illustrated below by using the glucose determination method of Beijing Leadman Biochemical Co., Ltd., using the method of the present invention and ultracentrifugation and adopting the glucose determination method of Leadman Biochemical Co., Ltd. to measure glucose respectively.

[0036] 1. The method of the present invention: on the OLYMPUS AU2700 fully automated biochemical analyzer in Japan, the instrument automatically adds 3 μl of sample to 225 μl of reagent I and mixes evenly, incubates at 37°C for 3 minu...

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Abstract

The invention discloses a method for measuring glucose oxidase without the interference of blood lipid. The method utilizes visible light to test a material according to the color change of reactions.According to the method, a reagent I comprises lipoprotein lipase, glycerol dehydrogenase, NAD+, Triton X-100, and mutarotase; and a reagent II comprises following effective components: glucose oxidase, peroxidase, and 4-amino antipyrine, and 2,4-dichlorophenol. The method comprises the following steps: placing serum and the reagent I in a water bath with a temperature of 37 DEG C for 3 to 5 minutes so as to convert triglyceride into dihydroxyl acetone under the action of lipoprotein lipase and glycerol dehydrogenase; and then adding the reagent II. Under the action of glucose oxidase and peroxidase, glucose, 4-amino antipyrine, and 2,4-dichlorophenol carry out condensation reactions to generate red quinone-imine. The first step is taken as blank by an instrument, and the glucose contentis calculated on the basis of quinone-imine generated in the step two.

Description

technical field [0001] The invention belongs to a method for measuring enzymes; or a method for testing materials by using visible light to produce color changes as a result of a test reaction, in particular to a method for detecting glucose in serum with a biochemical analyzer. Background technique [0002] The commonly used detection methods for serum glucose mainly include glucose oxidation method, microcurrent method, oxygen rate method, hexokinase method, o-toluidine method, glucose dehydrogenase method, gas chromatography / isotope dilution mass spectrometry, etc., which are recommended by domestic clinical laboratories Using the glucose oxidase method. Glucose oxidase (glucose oxidase, GOD) uses oxygen and water to oxidize glucose to gluconic acid and release hydrogen peroxide. Peroxidase (peroxidase, POD) dehydrocondenses hydrogen peroxide, 4-aminoantipyrine and phenol into red quinones, which is the Trinder reaction, and the amount of red quinones produced is proport...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N21/31
CPCG01N21/31G01N21/78
Inventor 李立和丁弘刘成旺
Owner TIANJIN BAODI HOSPITAL
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